9-amino-7-(substituted)-6-demethyl-6-deoxytetracyclines

ABSTRACT

9-amino-7-(substituted amino)-6-demethyl-6-deoxytetracyclines having activity against a wide spectrum of organisms including organisms which are resistant to tetracyclines are disclosed. Also disclosed are intermediates and methods for making the novel compounds of the present invention.

FIELD OF THE INVENTION

The invention relates to novel[4S-(4α,12α)]-9-amino-4-(dimethylamino)-7-(substitutedamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamides,hereinafter called 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines, which exhibit antibioticactivity against a wide spectrum of organisms including organisms whichare resistant to tetracyclines and are useful as antibacterial agents.

The invention also relates to novel 7-(substitutedamino)-9-nitro-6-demethyl-6-deoxytetracycline compounds useful formaking the novel compounds of the present invention and to novel methodsfor producing the novel compounds and intermediate compounds.

DESCRIPTION OF THE PRIOR ART

A variety of tetracycline antibiotics have been synthesized anddescribed for the treatment of infectious diseases in man and animalssince 1947. Tetracyclines inhibit protein synthesis by binding to the30S subunit of the bacterial ribosome preventing binding of aminoacylRNA (Chopra, Handbook of Experimental Pharmacology, Vol. 78, 317-392,Springer-Verlag, 1985). Resistance to tetracyclines has emerged amongmany clinically important microorganisms which limit the utility ofthese antibiotics. There are two major mechanisms of bacterialresistance to tetracyclines: a) energy-dependent efflux of theantibiotic mediated by proteins located in the cytoplasmic membranewhich prevents intracellular accumulation of tetracycline (S. B. Levy,et al., Antimicrob. Agents Chemotherapy 33, 1373-1374 (1989); and b)ribosomal protection mediated by a cytoplasmic protein which interactswith the ribosome such that tetracycline no longer binds or inhibitsprotein synthesis (A. A. Salyers, B. S. Speers and N. B. Shoemaker, Mol.Microbiol, 4:151-156, 1990). The efflux mechanism of resistance isencoded by resistance determinants designated tetA-tetL. They are commonin many Gram-negative bacteria (resistance genes Class A-E), such asEnterobacteriaceae, Pseudomonas, Haemoohilus and Aeromonas, and inGram-positive bacteria (resistance genes Class K and L), such asStaphylococcus, Bacillus and Streptococcus. The ribosomal protectionmechanism of resistance is encoded by resistance determinants designatedTetM, N and O, and is common in Staphylococcus, Streptococcus,Campylobacter, Gardnerella, Haemoohilus and Mycoolasma (A. A. Salyers,B. S. Speers and N. B. Shoemaker, Mol. Microbiol, 4:151-156 1990).

A particularly useful tetracycline compound is7-(dimethylamino)-6-demethyl-6-deoxytetracycline, known as minocycline(see U.S. Pat. Nos. 3,148,212, 26,253 and 3,226,436 discussed below).However, strains harboring the tetB (efflux in gram-negative bacteria)mechanism, but not tetK (efflux in Staphylococcus) are resistant tominocycline. Also, strains carrying tetM (ribosomal protection) areresistant to minocycline. This invention describes the synthesis ofnovel tetracycline compounds which demonstrate significant in vitro andin vivo activity vs. tetracycline and minocycline susceptible strainsand some tetracycline and minocycline resistant strains, that is, thoseharboring the tetM (ribosomal protection) resistance determinants.

Duggar, U.S. Pat. No. 2,482,055, discloses the preparation ofAureomycin® (I) by fermentation which have antibacterial activity.Growich et al., U.S. Pat. No. 3,007,965, disclose improvements to thefermentation preparation of I. Neither of these patents teaches orsuggests the 6-demethyl-6-deoxytetracyclines ##STR1## Beereboom et al.,U.S. Pat. No. 3,043,875 discloses tetracycline derivatives of theformulae (II) and (III) where R is H or CH₃ ; R₁ is H and when R is CH₃; OH; R₂ is H and N(CH₃)₂ ; X and Y are halogen; Z is H and halogen andB is bromo, chloro and iodo, which have antibacterial activity. Thispatent does not teach or suggest the inclusion of both di(loweralkyl)amino or mono(lower alkyl)amino substituents (at Y or Z) and anamino function (at B). ##STR2## Boothe et al., U.S. Pat. No. 3,148,212,reissued as U.S. Pat. No. 26,253, and Petisi et al., U.S. Pat. No.3,226,436, discloses tetracycline derivatives of the formula (IV)wherein R is hydrogen or methyl and R₁ and R₂ is hydrogen, mono(loweralkyl)amino or di(lower alkyl)amino with the proviso that R₁ and R₂cannot both be hydrogen, which are useful for treating bacterialinfections. This patent does not teach or suggest the inclusion of a9-amino functionality (at R₂). ##STR3## Blackwood et al., U.S. Pat. No.3,200,149 discloses tetracycline derivatives of the formulae (V) and(VI) and reduction products thereof wherein Y may be hydrogen orhydroxyl, X may be hydrogen, chloro, iodo, or bromo, X₁ may be hydrogen,amino, and lower alkanoylamino, X₂ may be hydrogen or nitro and Z ischloro or fluoro which possess microbiological activity. This patentdoes not teach or suggest the inclusion of both a di(lower alkyl)aminogroup (at X) and another nitrogen functionality (at X₁) on the6-demethyl-6-deoxytetracycline nucleus. ##STR4## Petisi et al., U.S.Pat. No. 3,338,963 discloses tetracycline compounds of the formula (VII)wherein R₁ and R₂ are hydrogen, nitro, amino, formylamino, acetylamino,p-(dihydroxyboryl)benzoylamino, p-(aminobenzenesulfonyl)amino, chlorine,bromine or diazonium with the proviso that R₁ and R₂ may not both behydrogen and with the further proviso that when R₁ is chlorine orbromine, R may not be hydrogen and vice versa, R₃ is hydrogen or methyland R₄ is hydrogen or hydroxy, which have broad-spectrum antibacterialactivity. This patent does not teach or suggest the inclusion of bothdi(lower alkyl)amino or mono(lower alkyl)amino substituents (at R₁ ) andamino substituents (at R₂). ##STR5## Bitha et al., U.S. Pat. No.3,341,585 discloses tetracycline compounds of the formula (VIII) whereinR₅ is hydrogen, α-hydroxy or α-hydroxy, R₆ is α-methyl or α-methyl, andR₇ and R₉ are each hydrogen, mono(lower alkyl)amino or di(loweralkyl)amino with the proviso that R₇ and R₉ cannot both be hydrogen andwith the further proviso that when R₅ is hydrogen then R₆ is β-methyl. Apreferred embodiment of the general formula (VIII) is when R₅ isα-hydroxy or β-hydroxy, R₆ is α-methyl or β-methyl, R₇ is di(loweralkyl)amino and R₉ is hydrogen, which have broad-spectrum antibacterialactivity This patent does not teach or suggest the inclusion of bothdi(lower alkyl)amino or mono(lower alkyl)amino substituents (at R₇) andamino substituents (at R₉). ##STR6## Shu, U.S. Pat. No. 3,360,557discloses 9-hydroxytetracyclines of the formula (IX) wherein R₁ ishydrogen or hydroxy, R₂ is hydrogen or hydroxy, R₃ is hydrogen ormethyl, R₂ and R₃ taken together is methylene, and R₄ is hydrogen,halogen, nitro, amino, mono(lower alkyl)amino or di(lower alkyl)amino,which have been found to possess antibacterial activity. This patent isrestricted to 9-hydroxytetracyclines and does not teach or suggest thepresently claimed compounds. ##STR7## Zambrano, U.S. Pat. No. 3,360,561discloses a process for preparing 9-nitrotetracyclines of the formula(X) wherein R₅ is hydrogen or hydroxy, R₁ is hydrogen or hydroxy, R₆ ishydrogen or methyl, R₁ and R₆ taken together is methylene, R₇ ishydrogen, chloro or nitro and R₉ is hydrogen or nitro with the provisothat R₇ and R₉ cannot both be hydrogen. This patent does not teach orsuggest the inclusion of both a di(lower alkyl)amino or mono(loweralkyl)amino substituent (at R₇) and an amino functionality (at R₉).##STR8## Martell et al., U.S. Pat. No. 3,518,306 discloses 7-and/or9-(N-nitrosoalkylamino)-6-demethyl-6-deoxytetracyclines of the formula(XI) which possess in vivo antibacterial activity. This patent does notteach or suggest the inclusion of both a di(lower alkyl)amino ormono(lower alkyl)amino substituent (at C-7) and an amino functionality(at C-9). ##STR9##

In U.S. Pat. No. 5,021,407, a method of overcoming the resistance oftetracycline resistant bacteria is disclosed. The method involvesutilizing a blocking agent compound in conjunction with a tetracyclinetype antibiotic. This patent does not disclose novel tetracyclinecompounds which themselves have activity against resistant organisms.

In summary, none of the above patents teach or suggest the novelcompounds of this application. In addition, none of the above patentsteach or suggest novel tetracycline compounds having activity againsttetracycline and minocycline resistant strains as well as strains whichare normally susceptible to tetracyclines.

SUMMARY OF THE INVENTION

This invention is concerned with novel 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines, represented by formula I and II,which have antibacterial activity, with methods of treating infectiousdiseases in warm-blooded animals employing these new compounds; withmethods of treating or controlling veterinary diseases; withpharmaceutical preparations containing these compounds; with novelintermediate compounds and processes for the production of thesecompounds. More particularly, this invention is concerned with compoundsof formula I which have enhanced in vitro and in vivo antibacterialactivity against tetracycline resistant strains as well as a high levelof activity against strains which are normally susceptible totetracyclines. ##STR10## In formula I and II, R=NR₁ R₂, and when R₁=hydrogen, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl; and when R₁ =methylor ethyl, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =n-propyl, R₂ =n-propyl,1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylethyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and whenR₁ =n-butyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylpropyl, R₂ =2-methylpropyl; R₃ is selected from hydrogen,straight or branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propylor 1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl, α-naphthyl orβ-naphthyl; (C₇ --C₉)aralkyl group such as benzyl, 1-phenylethyl,2-phenylethyl or phenylpropyl; a heterocycle group such as 2 or3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substitutedpyridyl, benzofuranyl, benzothienyl, quinolinyl or --(CH₂).sub. nCOOR₅when n=1-4 and R₅ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆-C₁₀) aryl group such as phenyl, α-naphthyl, β-naphthyl; R₄ is selectedfrom hydrogen; straight or branched (C₁ -C₃)alkyl group such as methyl,ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl,α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl,1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocycle group suchas 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl such as methyl, ethyl, n-propyl or1-methylethyl; or (C₆ -C₁₀)aryl such as phenyl, α-naphthyl orβ-naphthyl; or R₃ and R₄ taken together are --(CH₂)₂ W(CH₂)-- wherein Wis selected from (CH₂)_(n) and n=0-1, --NH, --N(C₁ -C₃)alkyl, --N(C₁-C₄)alkoxy, oxygen, sulfur or substituted congeners selected from (L orD) proline, ethyl (L or D) prolinate, morpholine, pyrrolidine orpiperidine; and the pharmacologically acceptable organic and inorganicsalts, and metal complexes.

Particularly preferred are compounds according to the above formula Iand II in which R=NR₁ R₂, and when R₁ =hydrogen, R₂ =ethyl, n-propyl,1-methylethyl, n-butyl, 1-methylpropyl, 2-methylpropyl or1,1-dimethylethyl; and when R₁ =methyl, R₂ =methyl, ethyl, n-propyl,n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁ =ethyl, R₂=ethyl, n-propyl, n-butyl or 2-methylpropyl; and when R₁ =n-propyl, R₂=n-propyl, n-butyl or 2-methylpropyl; and when R₁ =n-butyl, R₂ =n-butylor 2-methylpropyl; R₃ is selected from hydrogen, straight or branched(C₁ -C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl;(C₆ -C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇-C₉)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl; a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or --(CH₂)_(n) COOR₅ when n=1-4and R₅ is selected from hydrogen; straight or branched (C₁ -C₃)alkylgroup such as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl group such as phenyl, α-naphthyl, β-naphthyl; R₄ is selected fromhydrogen; straight or branched (C₁ -C₃)alkyl group such as methyl,ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl,α-naphthyl or β-naphthyl; (C₇ -C₉) aralkyl group such as benzyl,1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocycle group suchas 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl such as methyl, ethyl, n-propyl or1-methylethyl; or )aryl such as phenyl, α-naphthyl or β-naphthyl; or R₃and R₄ taken together are --(CH₂)₂ W(CH₂)--, wherein W is selected from(CH₂)_(n) and n=0-1, --NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen,sulfur or substituted congeners selected from (L or D) proline, ethyl (Lor D) prolinate, morpholine, pyrrolidine or piperidine; and thepharmacologically acceptable organic and inorganic salts, and metalcomplexes.

Most particularly preferred are compounds according to the above formulaI and II in which R=NR₁ R₂, and when R₁ =hydrogen, R₂ =ethyl, n-propylor 1-methylethyl; and when R₁ =methyl, R₂ =methyl, ethyl or n-propyl;and when R₁ =ethyl, R₂ =ethyl; R₃ is selected from hydrogen, straight orbranched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propyl or1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl, α-naphthyl orβ-naphthyl; (C₇ -C₉)aralkyl group such as benzyl, 1-phenylethyl,2-phenylethyl or phenylpropyl; a heterocycle group such as 2 or3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substitutedpyridyl, benzofuranyl, benzothienyl, quinolinyl or --(CH₂ _(nCOOR) ₅when n=1-4 and R₅ is selected from hydrogen; straight or branched (Calkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆-C₁₀ ) aryl group such as phenyl, α-naphthyl, β-naphthyl; R₄ is selectedfrom hydrogen; straight or branched (C₁ -C₃)alkyl group such as methyl,ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl,α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl,1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocycle group suchas 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C )alkyl such as methyl, ethyl, n-propyl or 1-methylethyl:or (C₆ -C₁₀)aryl such as phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄taken together are --(CH₂)₂ W(CH₂)--, wherein W is selected from(CH₂)_(n) and n=0-1, --NH, --N(C₁ - C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen,sulfur or substituted congeners selected from (L or D) proline, ethyl (Lor D) prolinate, morpholine, pyrrolidine or piperidine; and thepharmacologically acceptable organic and inorganic salts, and metalcomplexes.

Also included in the present invention are compounds useful asintermediate for producing the above compounds of formula I. Suchintermediate compounds include those having the formulae: ##STR11##wherein: R=NR₁ R₂, and when R₁ =hydrogen, R₂, =methyl, ethyl, n-propyl,1-methylethyl, n-butyl, 1-methylpropyl, 2-methylpropyl or1,1-dimethylethyl; and when R₁ =methyl or ethyl, R₂ =methyl, ethyl,n-propyl, 1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl; andwhen R₁ =n-propyl, R₂ =n-propyl, 1-methylethyl, n-butyl, 1-methylpropylor 2-methylpropyl; and when R₁ =1-methylethyl, R₂ =n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =n-butyl, R₂ =n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =1-methylpropyl, R₂=2-methylpropyl; R₃ is selected from hydrogen, straight or branched (₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; (C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇ C₉)aralkylgroup such as benzyl, 1-phenylethyl, 2-phenylethyl or phenylpropyl; aheterocycle group such as 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl, benzofuranyl,benzothienyl, quinolinyl or --(CH₂ _(n) COOR₅ when n=1-4 and R₅ isselected from hydrogen; straight or branched (C₁ -C₃)alkyl group such asmethyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀) aryl group suchas phenyl, α-naphthyl, β-naphthyl; R₄ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propylor 1-methylethyl; (C₇ -C₉)aryl group such as phenyl, α-naphthyl orβ-naphthyl; (C₇ -C₉)aralkyl group such as benzyl, 1-phenylethyl,2-phenylethyl or phenylpropyl; a heterocycle group such as 2 or3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substitutedpyridyl, benzofuranyl, benzothienyl, quinolinyl or --(CH₂)_(nCOOR) ₆when n=1-4 and R₆ is selected from hydrogen; straight or branched (C₁-C₃)alkyl such as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆-C₁₀)aryl such as phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄ takentogether are --(CH₂)₂ W(CH₂)--, wherein W is selected from (CH₂)_(n) andn=0-1, --NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur orsubstituted congeners selected from (L or D) proline, ethyl (L or D)prolinate, morpholine, pyrrolidine or piperidine; and thepharmacologically acceptable organic and inorganic salts, and metalcomplexes.

Particularly preferred are compounds according to the above formula IIIand IV in which R=NR₁ R₂, and when R₁ =hydrogen, R₂ =ethyl, n-propyl,1-methylethyl, n-butyl, 1-methylpropyl, 2-methylpropyl or1,1-dimethylethyl; and when R₁ =methyl, R₂ =methyl, ethyl, n-propyl,n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁ =ethyl, R₂=ethyl, n-propyl, n-butyl or 2-methylpropyl; and when R₁ =n-propyl, R₂=n-propyl, n-butyl or 2-methylpropyl; and when R₁ =n-butyl, R₂ =n-butylor 2-methylpropyl; R₃ is selected from hydrogen, straight or branched (₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; (C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇-C₉)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl; a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ - C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or --(CH₂)_(n) COOR₅ when n=1-4and R₅ is selected from hydrogen; straight or branched (C₁ -C₃)alkylgroup such as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl group such as phenyl, α-naphthyl, β-naphthyl; R₄ is selected fromhydrogen; straight or branched (C₁ -C₃)alkyl group such as methyl,ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl,α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl,1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocycle group suchas 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl such as methyl, ethyl, n-propyl or1-methylethyl; or (C₆ -C₁₀)aryl such as phenyl, α-naphthyl orβ-naphthyl; or R₃ and R₄ taken together are --(CH₂)₂ W(CH₂)--, wherein Wis selected from (CH₂)_(n) and n=0-1, --NH, --N(C )alkyl, --N(C₁-C₄)alkoxy, oxygen, sulfur or substituted congeners selected from (L orD) proline, ethyl (L or D) prolinate, morpholine, pyrrolidine orpiperidine; and the pharmacologically acceptable organic and inorganicsalts, and metal complexes.

Most particularly preferred are compounds according to the above formulaIII and IV in which R=NR₁,R₂, and when R₁ =hydrogen, R₂ =ethyl, n-propylor 1-methylethyl; and when R₁ =methyl, R₂ =methyl, ethyl or n-propyl;and when R₁ =ethyl; R₂ =ethyl; R₃ is selected from hydrogen, straight orbranched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propyl or1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl, α-naphthyl orβ-naphthyl; (C₇ -C₉)aralkyl group such as benzyl, 1-phenylethyl,2-phenylethyl or phenylpropyl; a heterocycle group such as 2 or3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substitutedpyridyl, benzofuranyl, benzothienyl, quinolinyl or --(CH₂)_(n) COOR₅when n=1-4 and R₅ is selected from hydrogen; straight or branched (C₁C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆-C₁₀) aryl group such as phenyl, α-naphthyl, β-naphthyl; R₄ is selectedfrom hydrogen; straight or branched (C₁ -C₃)alkyl group such as methyl,ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl group such as phenyl,α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl,1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocycle group suchas 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl such as methyl, ethyl, n-propyl or1-methylethyl; or (C₆ -C₁₀)aryl such as phenyl, α-naphthyl orβ-naphthyl; or R₃ and R₄ taken together are --(CH₂ ₂ W(CH₂)--, wherein Wis selected from (CH₂)_(n) and n=0-1, --NH, --N(C₁ -C₃)-- alkyl, --N(C₁-C₄)alkoxy, oxygen, sulfur or substituted congeners selected from (L orD) proline, ethyl (L or D) prolinate, morpholine, pyrrolidine orpiperidine; and the pharmacologically acceptable organic and inorganicsalts, and metal complexes.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The novel compounds of the present invention may be readily prepared inaccordance with the following schemes.

The starting 7-(substituted amino)-6-demethyl-6-deoxytetracyclinesdescribed in formula 1, wherein R=NR₁ R₂ and R₁ =R₂ (1a) and R=NHR₂ (1b)or the salts thereof are prepared by procedures known to those skilledin the art including those described in U.S. Pat. Nos. 3,226,436 and3,518,306. ##STR12##

The starting 7-(substituted amino)-6-demethyl-6-deoxytetracyclinesdescribed in formula 1 wherein R=NR₁ R₂ and R₁ ≠R₂ (1c) are preparedaccording to Scheme 1. ##STR13##

In accordance with Scheme I, a7-(monoalkylamino)-6-demethyl-6-deoxytetracycline, 1b, in which R=NHR₂,is reductively alkylated with an aldehyde to give an unsymmetricaldialkylamino, 1c. ##STR14##

In accordance with Scheme II, a 7-(substitutedamino)-6-demethyl-6-deoxytetracycline or its salts, 1a-1c, is treatedwith

a) a metal nitrate salt; such as calcium, potassium or sodium; and astrong acid; such as sulfuric acid, trifluoroacetic acid,methanesulfonic acid or perchloric acid or

b) nitric acid and a strong acid; such as sulfuric acid, trifluoroaceticacid, methanesulfonic acid or perchloric acid; to form the corresponding7-(substituted amino)-9-nitro-6-demethyl-6-deoxytetracycline 2.

To produce the 9-(amino)-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines of the present invention,compound 2 or its salts is treated with hydrogen in an acid alcoholsolvent, preferably 2-methoxyethanol, in the presence of a suitablecatalyst such as, for example: a) any supported catalyst; such as0.5-25% palladium-on-carbon, 0.5-25%- palladium-on-barium sulfate,0.5-25% platinum-on-carbon or 0.5-25% rhodium-on-carbon;

b) any reducible metal oxide catalyst; such as Raney nickel or platinumoxide; or

c) a homogeneous hydrogenation catalyst; such astris(triphenylphosphine)rhodium (I) chloride; to obtain the9-amino-7-(substituted amino)-6-demethyl-6-deoxytetracycline, 3.

Alternatively, the 9-(amino)-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines of the present invention areobtained by treating with:

a) stannous chloride dihydrate as described by R. B. Wordward, Org.Syn., Coll. Vol. 3, 453 (1955);

b) a soluble metal sulfide, preferably sodium sulfide, in alcoholicsolvents as described by G. R. Robertson, Org. Syn., Coll. Vol. 1, 52(1941);

c) an active metal in mineral acid; such as iron, tin or zinc in dilutehydrochloric acid;

d) active metal couples; such as copper-zinc, tin-mercury or aluminumamalgam in dilute acid; or

e) transfer hydrogenation using triethylammonium formate and a supportedcatalyst as described by I. D. Entwistle et al., J. Chem. Soc., Perkin1, 443 (1977). ##STR15##

In accordance with Scheme III, Z=NO₂ or NH₂ ; compound 4 is selectivelyN-alkylated in the presence of formaldehyde and either a primary aminesuch as methylamine, ethylamine, benzylamine, methyl glycinate, (L or D)lysine, (L or D) alanine or their substituted congeners; or a secondaryamine such as morpholine, pyrrolidine, piperidine or their substitutedcongeners to give the corresponding Mannich base adducts, 5, of thebiologically active 7-(substitutedamino)-6-demethyl-6-deoxytetracyclines. Contemplated equivalents includethose substituted morpholine, pyrrolidine or piperidine moieties whereinthe substituents are chosen to provide the requisite increase insolubility without adversely affecting antibacterial activity.

The 9-amino-7-(substituted amino)-6-demethyl-6-deoxytetracyclines, 3,may also be obtained as metal complexes such as aluminum, calcium, iron,magnesium, manganese and complex salts; inorganic and organic salts andcorresponding Mannich base adducts using methods known to those skilledin the art. Preferably, the 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines, are obtained as inorganic saltssuch as hydrochloric, hydrobromic, hydroiodic, phosphoric, nitric orsulfate; or organic salts such as acetate, benzoate, citrate, cysteineor other amino acids, fumarate, glycolate, maleate, succinate, tartrate,alkylsulfonate or arylsulfonate. In all cases, the salt formation occursWith the C(4)-dimethylamino group. The salts are preferred for oral andparenteral administration.

Biological Activity Methods for In Vitro Antibacterial Evaluation (Table1)

The minimum inhibitory concentration (MIC), the lowest concentration ofthe antibiotic which inhibits growth of the test organism, is determinedby the microtiter broth dilution method using 0.1 ml Muller-Hinton IIbroth (Baltimore Biological Laboratories) per well. A suitable oxygenscavenger (i.e., cysteine or dithiothreitol) is added to the assaymedium for the testing of compounds according to Formula I because ofthe sensitivity of those compounds to oxidation. An inoculum level of1-5×10⁵ CFU/ml and a range of antibiotic concentrations (32 0.004 μg/ml)are used. MIC's were determined after the plates were incubated for 18hours at 35° C. in a forced air incubator.

E. coli in vitro protein translation system (Table 2)

The E. coli in vitro translation system can be used to study not onlythe mechanism of protein translation itself, but also the effect thatvarious compounds may have on protein synthesis. The system can be setup to function as a coupled transcription and translation system or as atranslation only system depending on whether DNA or RNA is added toinitiate protein synthesis. In this way, compounds affecting either RNAsynthesis and/or protein synthesis can be studied. Protein synthesis ismonitored by the incorporation of radiolabeled amino acids intotrichloroacetic acid precipitable material. The system used is basedupon literature methods [G. Zubay, Ann. Rev. Genet., 7: 267-287(1973)and J. Collins, Gene, 6: 28-42 (1979)].

The system used to study tetracycline protein synthesis inhibition is asfollows:

An S30 extract (supernatant from a 30,000×G centrifugation of lysedcells) of either tetracycline sensitive or tetracycline resistant(tetM+) cells is combined with a mixture of low molecular weightcompounds required for protein synthesis which include a mixture of 19amino acids, methionine, ³⁵ S-methionine, plasmid template DNA andeither dimethylsulfoxide (DMSO) or the tetracycline to be testeddissolved and diluted in DMSO. This mixture is incubated at 37° C. for30 minutes. Following the incubation, 2.5 μl of the 10 μl reaction isremoved and added to 0.5 ml of 1N sodium hydroxide. The solution isincubated an additional 15 minutes at 37° C., to destroy any m-RNA andt-RNA. The incorporation of ³⁵ S-methionine is determined byprecipitating the high molecular weight material in the sodium hydroxidealiquot with trichloroacetic acid (TCA), collecting the precipitatedmaterial on Whatman G/FC filters, drying the filters and counting theradioactivity retained on the filter. Percent inhibition (P.I.) ofprotein synthesis is determined by the following equation: ##EQU1##

In Vivo Antibacterial Evaluation (Table 3)

The therapeutic effects of tetracyclines are determined against acutelethal infections with various staphylococcal and E. coli strains.Female mice, strain CD-1 Charles River Laboratories, (20±2 grams) arechallenged by an intraperitoneal injection of sufficient bacteria(suspended in broth or hog gastric mucin) to kill non-treated controlswithin 24-48 hours. Antibacterial agents, contained in 0.5 ml of 0.2%aqueous agar, are administered subcutaneously or orally 30 minutes afterinfection. When an oral dosing schedule is used, animals are deprived offood for 5 hours before and 2 hours after infection. Five mice aretreated at each dose level. The 7 day survival ratios from threeseparate tests are pooled for calculation of median effective dose(ED₅₀).

Legend for Tables I-III

A=9-Amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride

B=7-(Dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(minocycline hydrochloride)

C=9-Amino-7-(diethylamino)-6-demethyl-6-deoxytetracycline sulfate

D=7-(Diethylamino)-6-demethyl-6-deoxytetracycline sulfate

                  TABLE 1                                                         ______________________________________                                        In Vitro Antibacterial Activity of                                            6-Demethyl-6-deoxytetracycline Derivatives                                               MIC (ug/ml)                                                        Organism*    A.sup.a  B        C.sup.a                                                                              D                                       ______________________________________                                        S. aureus UBMS 88-5                                                                        0.06     4        0.5    16                                      (tetM)                                                                        S. aureus UBMS 88-4                                                                        0.015    0.008    0.03   0.03                                    (tetracycline-sensitive)                                                      S. aureus UBMS 90-1                                                                        0.25     4        2      8                                       (tetM)                                                                        S. aureus UBMS 90-2                                                                        0.06     1        0.25   8                                       (tetM)                                                                        S. aureus UBMS 90-3                                                                        0.015    0.015    0.03   0.03                                    (tetracycline-sensitive)                                                      S. aureus UBMS 88-7                                                                        0.12     0.03     0.06   0.12                                    (tetK)                                                                        S. aureus IVES 2943                                                                        0.5      1        0.25   8                                       (methicillin-resistant)                                                       S. aureus IVES 1983                                                                        0.5      1        0.25   8                                       (methicillin-resistant)                                                       S. aureus CI 2371                                                                          0.5      4        NA     NA                                      (methicillin-resistant)                                                       S. aureus CI 3300                                                                          0.25     8        NA     NA                                      (methicillin-resistant)                                                       Coagulase negative                                                                         0.003    0.015    NA     NA                                      staphylococci CI 664                                                          Coagulase negative                                                                         1        8        NA     NA                                      staphylococci CI 535                                                          S. haemolyticus                                                                            0.06     0.12     0.12   NA                                      AVAH 88-3                                                                     E. faecalis AMV 120                                                                        4        16       NA     NA                                      (tetM)                                                                        E. faecalis PAM 211                                                                        4        16       NA     NA                                      (tetN)                                                                        E. faecalis 12201                                                                          0.5      4        NA     NA                                      (vancomycin-resistant)                                                        E. faecalis CI 2735                                                                        0.5      4        NA     NA                                      E. coli UBMS 88-1                                                                          >32      8        2      32                                      (tetB)                                                                        E. coli UBMS 88-2                                                                          0.25     0.5      0.5    2                                       (tetracycline-sensitive)                                                      E. coli UBMS 89-1                                                                          1        8        2      NA                                      (tetM)                                                                        E. coli UBMS 89-2                                                                          0.5      0.5      0.5    4                                       (tetracycline-sensitive)                                                      E. coli UBMS 90-4                                                                          4        >32      32     >32                                     (tetM)                                                                        E. coli UBMS 90-5                                                                          0.25     0.5      1      2                                       (tetracycline-sensitive)                                                      M. morganii NEMC                                                                           2        2        2      32                                      87-119                                                                        S. marcescens FPOR                                                                         4        2        4      32                                      87-33                                                                         P. aeruginosa ATCC                                                                         2        4        8      32                                      27853                                                                         X. maltophilia FPOR                                                                        0.12     0.06     0.12   0.25                                    87-210                                                                        E. coli ATCC 25922                                                                         0.25     0.25     0.50   1                                       E. faecalis ATCC                                                                           0.06     0.25     0.12   8                                       29212                                                                         S. aureus ATCC 29213                                                                       0.008    ≦0.004                                                                          ≦0.015                                                                        <0.015                                  ______________________________________                                         .sup.a In vitro assay is done in the presence of cysteine (0.05%).            Antibacterial potency of B and D was not enhanced in the presence of          cysteine.                                                                     *The tetM resistance determinant protects ribosomes from tetracycline, th     tetK determinant promotes efflux of the drug from the cell.              

                                      TABLE 2                                     __________________________________________________________________________    In Vitro Protein Translation with E. coli S30 Ribosomes                       __________________________________________________________________________    Wild Type Version                                                                     B                     A                                                       Wild Type  Wild Type  Wild Type  Wild Type                                    S30 no DTT*                                                                              S30 with DTT*                                                                            S30 no DTT*                                                                              S30 with DTT                         Reaction                                                                              Counts                                                                            % Inhibition                                                                         Counts                                                                            % Inhibition                                                                         Counts                                                                            % Inhibition                                                                         Counts                                                                            % Inhibition                     __________________________________________________________________________    Control DMSO                                                                          457268     872132     457268     872132                               Plus compound                                                                 1.0 mg/ml                                                                              57696                                                                            87      58885                                                                            93      52595                                                                            88      38121                                                                            96                               1:2 Dilution                                                                           76804                                                                            83      78961                                                                            91      80494                                                                            82      60891                                                                            93                               1:4 Dilution                                                                           53400                                                                            88     111971                                                                            87      95015                                                                            79      92203                                                                            89                               1:8 Dilution                                                                          162405                                                                            64     149792                                                                            83     130209                                                                            72     143215                                                                            84                               1:16 Dilution                                                                         213077                                                                            53     207484                                                                            76     205392                                                                            55     214831                                                                            75                               1:32 Dilution                                                                         306650                                                                            33     289304                                                                            67     297786                                                                            35     238321                                                                            73                               1:64 Dilution                                                                         457601                                                                            0      518601                                                                            41     245494                                                                            46     380103                                                                            56                               __________________________________________________________________________    tetM Variant                                                                          tetM S30 no DTT*                                                                         tetM S30 with DTT*                                                                       tetM S30 no DTT*                                                                          tetM S30 with DTT*                  Reaction                                                                              Counts                                                                            % Inhibition                                                                         Counts                                                                            % Inhibition                                                                         Counts                                                                            % Inhibition                                                                         Counts                                                                            % Inhibition                     __________________________________________________________________________    Control DMSO                                                                          247938     447247     247938     447247                               Plus compound                                                                 1.0 mg/ml                                                                             281528                                                                            0      371475                                                                            17     129780                                                                            48     145068                                                                            68                               1:2 Dilution                                                                          258633                                                                            0      331439                                                                            26     158861                                                                            36     168574                                                                            62                               1:4 Dilution                                                                          248904                                                                            0      373168                                                                            17     203184                                                                            18     226708                                                                            49                               1:8 Dilution                                                                          289595                                                                            0      421533                                                                             6     231447                                                                             7     284606                                                                            36                               1:16 Dilution                                                                         287498                                                                            0      493679                                                                             0     305633                                                                             0     375989                                                                            16                               1:32 Dilution                                                                         262329                                                                            0      490283                                                                             0     349994                                                                             0     412967                                                                             8                               1:64 Dilution                                                                         249242                                                                            0      452837                                                                             0     310723                                                                             0     507461                                                                             0                               __________________________________________________________________________     *Dithiothreitol (DTT) is used as an exygen scavenger.                    

                  TABLE 3                                                         ______________________________________                                        Effects of Compounds A and B on Acute                                         Lethal Infections in Mice                                                                Route of Antibiotic                                                                       ED.sub.50 (mg/kg).sup.+                                Organism     Administration*                                                                             A       B                                          ______________________________________                                        E. coli 311 (sens)                                                                         Subcutaneous  2.8     3.1                                        E. coli UBMS 90-4                                                                          Subcutaneous  24      >256                                       (tetM)                                                                        S. aureus UBMS 90-1                                                                        Subcutaneous  0.30    1.7                                        (tetM)                                                                        S. aureus UBMS 90-2                                                                        Oral          1.6     1.8                                        (tetM)       Subcutaneous  0.53    1.8                                        S. aureus Smith (sens)                                                                     Oral          0.81    0.53                                                    Subcutaneous  0.34    0.32                                       ______________________________________                                         *Single Dose                                                                  .sup.+ Median effective dose protecting 50% of the infected mice         

Testing Results

As seen from the above testing, the compounds according to the presentinvention display good activity against a spectrum of tetracyclinesensitive and resistant Gram-positive and Gram-negative bacteria,especially strains of E. coli, S. aureus and E. faecalis containing thetetM or tetK resistant determinants. As illustrated in Table I,9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(A) shows good in vitro activity against tetracycline resistant strainscarrying the tetM resistance determinant such as S. aureus UBMS 88-5, S.aureus UBMS 90-1 and 90-2, E. coli UBMS 89-1 and 90-4; and is equally aseffective as 7-(dimethylamino)-6-demethyl-6-deoxytetracyclinehydrochloride (B) vs. susceptible strains.7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride (B) and9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(A) are assayed in vitro for their ability to inhibit protein synthesis,taking place on either wild type or tetM protected ribosomes, using acoupled transcription and translation system. Similarly,9-amino-7-(diethylamino)-6-demethyl-6-deoxytetracycline sulfate (C)shows enhancement of antibacterial activity versus7-(diethylamino)-6-demethyl-6-deoxytetracycline sulfate (D).

Both compounds (A & B) are found to effectively inhibit proteinsynthesis on wild type ribosomes, having equivalent levels of activity(Table II). 7-(dimethylamino)-6-demethyl-6-deoxytetracyclinehydrochloride (B) is unable to inhibit protein synthesis occurring ontetM protected ribosomes. In contrast,9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(A) is effective in inhibiting protein synthesis occurring on tetMprotected ribosomes, although higher drug levels are required to achievesimilar levels of inhibition relative to wild type ribosomes.

The enhanced activity of9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline sulfate (A)against tetracycline susceptible and resistant organisms (tetM) isdemonstrated in Table 3 for animals infected with representativebacteria. Lowered ED₅₀ 's are obtained with9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline sulfate (A)than with 7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(B) in mice with of S. aureus and E. coli which carry the tetMresistance determinant. Similar ED₅₀ 's are obtained with9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline sulfate (A) and7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride (B)against infections with minocycline susceptible organisms.

As can be seen from Tables 1 and 3, the new 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines may be used to prevent or controlimportant veterinary diseases such as mastitis, diarrhea, urinary tractinfections, skin infections, ear infections, wound infections and thelike.

The improved efficacy of the new 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines is evidenced by the in vitroactivity against isogenic strains into which the resistant determinants,such as tetM were cloned (Table 1); the inhibition of protein synthesisby tetM resistant ribosomes (Table 2); and the in vivo activity againstexperimental infections caused by strains resistant to thetetracyclines, due to the presence of resistant determinants, tetM(Table 3).

When the compounds are employed as antibacterials, they can be combinedwith one or more pharmaceutically acceptable carriers, for example,solvents, diluents and the like, and may be administered orally in suchforms as tablets, capsules, dispersible powders, granules, orsuspensions containing, for example, from about 0.05 to 5% of suspendingagent, syrups containing, for example, from about 10 to 50% of sugar,and elixirs containing, for example, from about 20 to 50% ethanol, andthe like, or parenterally in the form of sterile injectable solutions orsuspensions containing from about 0.05 to 5% suspending agent in anisotonic medium. Such pharmaceutical preparations may contain, forexample, from about 25 to about 90% of the active ingredient incombination with the carrier, more usually between about 5% and 60% byweight.

An effective amount of compound from 2.0 mg/kg of body weight to 100.0mg/kg of body weight should be administered one to four times per dayvia any typical route of administration including but not limited tooral, parenteral (including subcutaneous, intravenous, intramuscular,intrasternal injection or infusion techniques), topical or rectal, indosage unit formulations containing conventional non-toxicpharmaceutically acceptable carriers, adjuvants and vehicles. It will beunderstood, however, that the specific dose level and frequency ofdosage for any particular patient may be varied and will depend upon avariety of factors including the activity of the specific compoundemployed, the metabolic stability and length of action of that compound,the age, body weight, general health, sex, diet, mode and time ofadministration, rate of excretion, drug combination, the severity of theparticular condition, and the host undergoing therapy.

These active compounds may be administered orally as well as byintravenous, intramuscular, or subcutaneous routes. Solid carriersinclude starch, lactose, dicalcium phosphate, microcrystallinecellulose, sucrose and kaolin, while liquid carriers include sterilewater, polyethylene glycols, non-ionic surfactants and edible oils suchas corn, peanut and sesame oils, as are appropriate to the nature of theactive ingredient and the particular form of administration desired.Adjuvants customarily employed in the preparation of pharmaceuticalcompositions may be advantageously included, such as flavoring agents,coloring agents, preserving agents, and antioxidants, for example,vitamin E, ascorbic acid, BHT and BHA.

The preferred pharmaceutical compositions from the standpoint of ease ofpreparation and administration are solid compositions, particularlytablets and hard-filled or liquid-filled capsules. Oral administrationof the compounds is preferred.

These active compounds may also be administered parenterally orintraperitoneally. Solutions or suspensions of these active compounds asa free base or pharmacologically acceptable salt can be prepared inwater suitably mixed with a surfactant such as hydroxypropylcellulose.Dispersions can also be prepared in glycerol, liquid polyethyleneglycols and mixtures thereof in oils. Under ordinary conditions ofstorage and use, these preparations contain a preservative to preventthe growth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (e.g., glycerol, propylene glycol and liquidpolyethylene glycol), suitable mixtures thereof, and vegetable oil.

The invention will be more fully described in conjunction with thefollowing specific examples which are not to be construed as limitingthe scope of the invention.

EXAMPLE 1 [4S-(4α,12aα)]-4,7-Bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

To a stirred ice bath cooled solution of 0.444 g of[4S-(4α,12aα)]-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride, prepared by the procedure described in U.S. Pat. No.3,226,436, dissolved in 15 ml of sulfuric acid is added 0.101 g ofsodium nitrate. The mixture is stirred in the cold for 45 minutesfollowed by the dropwise addition to 500 ml of diethyl ether. Theresulting solid is collected, washed with diethyl ether and dried togive 0.6 g of the desired product as a solid. MS(FAB): m/z 503(M+H) and601(M+H₂ SO₄ +H).

EXAMPLE 2[4S-(4α,12aα)1-7-(Diethylamino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:2)

To a stirred ice cooled solution of 0.660 g of[4S-(4α,12aα)]-7-(diethylamino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride, prepared by the procedure described in U.S. Pat. No.3,226,436, dissolved in 15 ml of sulfuric acid is added 0.151 g ofsodium nitrate. The mixture is stirred in the cold followed by dropwiseaddition to 500 ml of diethyl ether. The resulting solid is collected,washed with diethyl ether and dried to give 0.8 g of the desired productas a solid. MS(FAB): m/z 531(M+H) and 629(M+H₂ SO₄ +H).

EXAMPLE 3

[4S-(4α,12aα)1-9-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a.6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

A mixture of 2.0 g of product from Example 1 in 20 ml of2-methoxyethanol is stirred for 10 minutes and filtered. The filtrate isshaken, in a pressure bottle, with 1.0 g of 10% palladium-on-carbon and5 ml of 2N sulfuric acid, under 30 lbs. of hydrogen pressure, for 1hour. The reaction mixture is filtered and the filtrate concentrated invacuo to half volume. The solution is poured into 100 ml of diethylether, the solid collected, washed with diethyl ether and dried to give1.6 g of the desired product as a solid. MS(FAB): m/z 473(M+H).

EXAMPLE 4[4S-(4α,12α)]-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride (1:1)

A mixture of 20.0 g of product from Example 1 in 250 ml of2-methoxyethanol is stirred for 10 minutes and filtered. The filtrate isshaken, in a pressure bottle, with 10.0 g of 10% palladium-on-carbon and100 ml of 1N ethanolic hydrogen chloride, under 30 lbs. of hydrogenpressure, for 1 hour. The reaction mixture is filtered and the filtrateconcentrated in vacuo to half volume. The solution is poured into 1 L ofdiethyl ether, the solid collected, washed with diethyl ether and driedto give 16.0 g of the desired product as an oil. The oil is suspended in20 ml of distilled water, made acidic with 2.8 ml of 32% hydrochloricacid and decolorized with charcoal. The mixture is filtered throughdiatomaceous earth and made basic (pH 4.0) with concentrated ammoniumhydroxide. The solid is collected at 4° C., washed with pH 4 water anddried in vacuo to give 14.2 g of the desired product as a solid.

¹ H NMR (CD₃ SOCD₃): δ 4.19(s,1H,4-H) and 7.29(s,1H,8-H). MS(FAB): m/z473(M+H).

Analysis for C₂₃ H₂₈ N₄ O₇ HCl 6.7% H₂ O. Calc'd: C,50.64; H,6.10;N,10.27; Cl,6.50. Found: C,50.72; H,6.07; N,10.27; Cl,6.62.

EXAMPLE 5[4S-(4α12α)]-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidep-toluenesulfonate (1:1)

The title compound is prepared by the procedure of Example 4, using 20 gof product from Example 1, to give 16.0 g of the desired product as thefree base. The free base is suspended in 20 ml of distilled water, madeacidic with p-toluenesulfonic acid monohydrate and decolorized withcharcoal. The mixture is filtered through diatomaceous earth and madebasic (pH 4.0) with concentrated ammonium hydroxide. The solid iscollected at 4° C., washed with pH 4 water and dried in vacuo to give16.0 g of the desired product.

EXAMPLE 6[4S-(4α,12α)]-Amino-7-(diethylamino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,1212a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:2)

The title compound is prepared by the procedure of Example 3, using 2.1g of product from Example 2, to give 1.5 g of the desired product as asolid. ¹ H NMR (CD₃ SOCD₃) δ 4.25(s,1H,4-H) and 7.27(s,1H,8-H). MS(FAB):m/z 501(M+H) and 599(M+H₂ SO₄ +H).

EXAMPLE 7[4S-(4α,12aα)]4-(Dimethylamino)-7-(ethylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride (1:1)

A solution of 0.460 g of[4S-(4α,12aα)]-4-(dimethylamino]-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride, prepared by the procedure described in U.S. Pat. No.3,226,436, 0.5 ml of 97% formic acid and 0.75 ml of 40% aqueousformaldehyde is heated at reflux temperature for 2 hours. The reactionmixture is cooled, concentrated in vacuo to half volume and poured intodiethyl ether. The resulting solid is collected, washed with diethylether and dried to give 0.3 g of the desired product as a solid.

EXAMPLE 8[4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydro-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

The title compound is prepared by the procedure of Example 1, using 0.46g of product from Example 7 to give 0.5 g of the desired product as asolid.

EXAMPLE 9[4S-(4α,12aα)]-Amino-4-(dimethylamino)-7-(ethylmethylamino)-1,4,4a,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

The title compound is prepared by the procedure of Example 3, using 1.0g of product from Example 8, to give 0.8 g of the desired product as asolid.

EXAMPLE 10[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

The title compound is prepared by the procedure of Example 1, using 0.48g of[4S-(4α,12aα)]-4-(dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,-6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride, prepared by the procedure described in U.S. Pat. No.3,226,436, to give 0.5 g of the desired product as a solid.

EXAMPLE 11[4S-(4α,12aα)]-Amino-4-(dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

The title compound is prepared by the procedure of Example 3, using 2.1g of product from Example 10, to give 1.5 g of the desired product as asolid.

EXAMPLE 12[4S-(4α12aα)]-4-(Dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

The title compound is prepared by the procedure of Example 1, using 0.96g of[4S-(4α,12aα)]-4-(dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamideprepared by the procedure described in U.S. Pat. No. 3,226,436, to give0.9 g of the desired product as a solid.

EXAMPLE 13[4S-(4α,12aα)]-Amino-4-(dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1)

The title compound is prepared by the procedure of Example 3, using 1.0g of product from Example 12, to give 0.7 g of the desired product as asolid.

EXAMPLES 14-35

Substantially following the methods described in detail hereinabove inExamples 3 and 9, the compounds of this invention listed below inExamples 14-35 are prepared.

EXAMPLE 14

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(methylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 15

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(butylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 16

4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[1-methylpropylamino)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 17

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 18

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-(ethylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 19

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 20

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-(butylethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 21

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 22

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 23

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(dipropyl-amino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 24

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methyethyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 25

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(butylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 26

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 27

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 28

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 29

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 30

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 31

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(dibutylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 32

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methyl-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 33

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 34

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 35

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLES 36-39

Substantially following the methods described in detail hereinabove inExamples 3 and 11, the compounds of this invention listed below inExamples 36-39 are prepared.

EXAMPLE 36

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(propylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 37

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(butylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 38

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 39

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1,1-dimethylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLES 40-65

Substantially following the methods described in detail hereinabove inExamples 1 and 2, the compounds of this invention listed below inExamples 40-65 are prepared.

EXAMPLE 40

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(methylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 41

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(butylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 42

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropylamino)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 43

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 44

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 45

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 46

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(butylethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 47

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 48

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 49

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(dipropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 50

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate

EXAMPLE 51

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(butylpropyltetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 52

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 53

4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 54

4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 55

4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 56

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 57

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(dibutylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 58

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 59

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 60

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 61

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 62

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(propylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 63

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(butylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 64

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 65

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1,1-dimethylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

We claim:
 1. A compound of the formula: ##STR16## wherein: R=NR₁ R₂, andwhen R₁ =hydrogen, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl; and when R₁ =methylor ethyl, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =n-propyl, R₂ =n-propyl,1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylethyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and whenR₁ =n-butyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylethyl, R₂ =2-methylpropyl; R₃ is selected from hydrogen,straight or branched (C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇-C₉)aralkyl group; or ----(CH₂)_(n) COOR₅ when n=1-4 and R₅ is selectedfrom hydrogen; straight or branched (C₁ -C₃)alkyl group; or (C₆-C₁₀)aryl group; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇ -C₉)aralkyl group; or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl; or (C₆ -C₁₀)aryl; and the pharmacologicallyacceptable organic and inorganic salts and metal complexes.
 2. Thecompound according to claim 1, wherein: R=NR₁ R₂, and when R₁ =hydrogen,R₂ =ethyl, n-propyl, 1-methylethyl, n-butyl, 1-methylpropyl,2-methylpropyl or 1,1-dimethylethyl; and when R₁ =methyl, R₂ =methyl,ethyl, n-propyl, n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=ethyl, R₂ =ethyl, n-propyl, n-butyl, or 2-methylpropyl; and when R₁=n-propyl, R₂ =n-propyl, n-butyl or 2-methylpropyl; and when R₁=n-butyl, R₂ =n-butyl or 2-methylpropyl; R₃ is selected from straight orbranched (C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇ -C₉)aralkylgroup; or (CH₂)_(n) COOR₅ when n=1-4 and R₅ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group; or (C₆ -C₁₀)aryl group; R₄ isselected from straight or branched (C₁ -C₃)alkyl group; (C₆ -C₁₀)arylgroup; (C₇ -C₉)aralkyl group; or --(CH₂)_(n) COOR₆ when n=1-4 and R₆ isselected from hydrogen; straight or branched (C₁ -C₃)alkyl; or (C₆-C₁₀)-aryl; and the pharmacologically acceptable organic salts and metalcomplexes.
 3. The compound according to claim 1, wherein: R=NR₁ R₂, andwhen R₁ =hydrogen, R₂ =ethyl, n-propyl or 1-methylethyl; and when R₁=methyl; R₂ =methyl, ethyl or n-propyl; and when R₁ =ethyl; R₂ =ethyl;R₃ is selected from a straight or branched (C₁ -C₃)alkyl group; (C₆-C₁₀)aryl group; (C₇ -C₉)aralkyl group; or --(CH₂)_(n) COOR₅ when n=1-4and R₅ is selected from hydrogen; straight or branched (C₁ -C₃)alkylgroup; or (C₆ -C₁₀)aryl group; R₄ is selected from straight or branched(C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇ -C₉)aralkyl group; or--(CH₂)_(n) COOR₆ when n=1-4 R₆ is selected from hydrogen; straight orbranched (C₁ -C₃)alkyl; or (C₆ -C₁₀)aryl; and the pharmacologicalacceptable organic and inorganic salts and metal complexes.
 4. Acompound of the formula: ##STR17## wherein: R=NR₁ R₂, and when R₁=hydrogen, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl; and when R₁ =methylor ethyl, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =n-propyl, R₂ =n-propyl,1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylethyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and whenR_(=n-butyl), R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylpropyl, R₂ =2-methylpropyl; R₃ is selected from hydrogen,straight or branched (C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇-C₉)aralkyl group; or --(CH₂)n.sub. COOR₅ when n=1-4 and R₅ is selectedfrom hydrogen; straight or branched (C₁ -C₃)alkyl group; or (C₆-C₁₀)aryl group; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group; (D₆ -C₁₀)aryl group; (C₇ -C₉)aralkyl group; or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl; or (C₆ -C₁₀)aryl; and the pharmacologicallyacceptable organic and inorganic salts and metal complexes.
 5. Thecompound according to claim 4, wherein R=NR₁ R₂, and when R₁ =hydrogen,R₂ =ethyl, n-propyl, 1-methylethyl, n-butyl, 1-methylpropyl,2-methylpropyl or 1,1,-dimethylethyl; and when R₁ =methyl, R₂ =methyl,ethyl, n-propyl, n-butyl, 1-methylpropyl, or 2-methylpropyl; and when R₁=ethyl, R₂ =ethyl, n-propyl, n-butyl, or 2-methylpropyl; and when R₁=n-propyl, R₂ =n-propyl, n-butyl, or 2-methylpropyl; and when R₁=n-butyl, R₂ =n-butyl or 2-methylpropyl; R₃ is selected from straight orbranched (C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇ -C₉)aralkylgroup; or (CH₂)_(n) COOR₅ when n=1-4 and R₅ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group; or (C₆ -C₁₀)aryl group; R₄ isselected from straight or branched (C₁ -C₃)alkyl, group; (C₆ -C₁₀)arylgroup; (C₇ -C₉)aralkyl group; or --(CH₂)_(n) COOR₆ when n=1-4 and R₆ isselected from hydrogen; straight or branched (C₁ -C₃)alkyl; or (C₆-C₁₀)aryl; and the pharmacologically acceptable organic and inorganicsalts and metal complexes; (C₁ -C₃)alkyl; (C₆ -C₁₀)aryl; and thepharmacologically acceptable organic and inorganic salts and metalcomplexes.
 6. The compound according to claim 4, wherein: R=NR₁ R₂, andwhen R₁ =hydrogen, R₂ =ethyl, n-propyl or 1-methylethyl; and when R₁=methyl, R₂ =methyl, ethyl or n-propyl; and when R₁ =ethyl, R₂ =ethyl;R₃ is selected from a straight or branched (C₁ -C₃)alkyl group; (C₆-C₁₀)aryl group; (C₇ -C₉)aralkyl group; or --(CH₂)_(n) COOR₅ when n=1-4and R₅ is selected from hydrogen; straight or branched (C₁ -C₃)alkylgroup; or (C₆ -C₁₀)aryl group; R₄ is selected from straight or branched(C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇ -C₉)aralkyl group; or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl; or (C₆ -C₁₀)aryl; pharmacologicallyacceptable organic and inorganic salts and metal complexes.
 7. Thecompound according to claim 1 wherein said inorganic salts comprise:hydrochloric, hydrobromic, hydroiodic, phosphoric, nitric or sulfate. 8.The compound according to claim 1 wherein said organic salts comprise:acetate, benzoate, citrate, cysteine or other amino acids, fumarate,glycolate, maleate, succinate, tartrate, alkylsulfonate orarylsulfonate.
 9. The compound according to claim 1 wherein said metalcomplexes comprise: aluminum, calcium, iron, magnesium, manganese andcomplex salts.
 10. The compound according to claim 4 wherein saidinorganic salts comprise: hydrochloric, hydrobromic, hydroiodic,phosphoric, nitric or sulfate.
 11. The compound according to claim 4wherein said organic salts comprise: acetate, benzoate, citrate, cysteinor other amino acids, fumarate, glycolate, maleate, succinate, tartrate,alkylsulfonate or arylsulfonate.
 12. The compound according to claim 4wherein said metal complexes comprise: aluminum, calcium, iron,magnesium, manganese and complex salts.
 13. The compound according toclaim 1,[4S-(4α,12aα)]-9-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 14. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12α-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride (1:1).
 15. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidep-toluenesulfonate (1:1).
 16. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-7-(diethylamino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:2).
 17. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(ethylmethylamino)-1,4,4a,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 18. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 19. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 20. The compound according to claim 4,[4S-(4α,12aα)]-4,7-Bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 21. The compound according to claim 4,[4S-(4α,12aα)]-7-(Diethylamino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:2).
 22. The compound according to claim 4,[4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydro-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 23. The compound according to claim 4,[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)-amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 24. The compound according to claim 4,[4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:1).
 25. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide26. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-7-(diethylamino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide.27. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(ethylmethylamino)-1,4,4a,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide.28. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide.29. The compound according to claim 1,[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide.30. The compound according to claim 4,[4S-(4α,12aα)]-4,7-Bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide.31. The compound according to claim 4,[4S-(4α,12aα)]-7-(Diethylamino)-4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide.32. The compound according to claim 4,[4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydro-9-nitro-1,11-dioxo-2-naphthacenecarboxamide.33. The compound according to claim 4,[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide.34. The compound according to claim 4,[4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide.35. A method for the prevention, treatment or control of bacterialinfections in warm-blooded animals which comprises administering to saidanimal a pharmacologically effective amount of a compound according toclaim
 1. 36. A pharmaceutical composition of matter comprising acompound according to claim 1 in association with a pharmaceuticallyacceptable carrier.
 37. A veterinary composition which comprises apharmacologically effective amount of a compound according to claim 1and pharmaceutically acceptable carrier.
 38. A method for theprevention, treatment or control of bacterial infections in warm-bloodedanimals caused by bacteria having the TetM and TetK resistantdeterminants which comprises administering to said animal apharmacologically effective amount of a compound according to claim 1.39. An antibacterial pharmaceutical composition of matter comprising aneffective amount of a compound according to claim 1 and an effectiveamount of an antioxidant in association with a pharmaceuticallyacceptable carrier.
 40. The composition of claim 39 wherein saidantioxidant comprises: cysteine, cysteine hydrochloride, dithiothreitol,ascorbic acid, monothioglycerol or thioglycolic acid.